The use of tobacco products has empirically observed, disparate effects in inflammatory bowel disease (IBD) (Birrenbach & Bocker (2004) Inflamm. Bowel Dis. 10:848-859; Karban & Eliakim (2007) World J. Gastroenterol. 13:2150-2152; Regueiro, et al. (2005) Inflamm. Bowel Dis. 11:42-47). Crohn's disease (CD) (Dalziel (1913) Br. Med. J. ii:1068-1070; Crohn, et al. (1932) J. Amer. Med. Assoc. 99:1323-1328) is exacerbated by the use of tobacco products (Regueiro, et al. (2005) supra; Cosnes, et al. (1999) Aliment Pharmacol. Ther. 13:1403-1411; Lewis, et al. (2007) J. Med. Genet. 44:689-694). In contrast, the clinical course of ulcerative colitis (UC) is ameliorated by use of tobacco (Regueiro, et al. (2005) supra; Aldhous, et al. (2007) Am. J. Gastroenterol. 102:589-597; Beaugerie, et al. (2001) Am. J. Gastroenterol. 96:2113-2116), or nicotine (Pullan, et al. (1994) N. Engl. J. Med. 330:811-815; McGrath, et al. (2004) Cochrane Database Syst. Rev.:CD004722) one of tobacco's 4000 constituent molecules (Dube & Green (1982) 36th Tobacco Chemists Research Conference. Symposium on the Formation, Analysis and Composition of Tobacco Smoke, Raleigh N.C. pp. 42-102; Jenkins, et al. (2000) The Chemistry of Environmental Tobacco:Composition and Measurement; Eisenberg (ed.) Boca Raton Fla.:CRC Press). The mechanism(s) involved although intensively investigated (Aldhous, et al. (2009) PLoS ONE 4:e6285; Nielsen, et al. (2009) PLoS ONE 4:e6210) is not understood (Karban & Eliakim (2007) supra), but is assumed to be due to the most bioactive component of tobacco; nicotine (Karban & Eliakim (2007) supra; Aldhous, et al. (2009) supra). It is of note that in a prior study, involving a solitary MAP strain, pure nicotine inhibited MAP growth in culture (Naser, et al. (2001) Am. J. Gastroenterol. 96:3455-3457).
The etiology of CD and UC is (are) not known. Mycobacterium avium subspecies paratuberculosis (MAP), causes a chronic wasting diarrheal disease in cattle called Johne's disease (Johne & Frothingham (1895) Dtsch. Zeitschr. Tiermed., Vergl. Pathol. 21:438-454), that is evocative of CD. Humans are continually exposed to viable MAP (Mishina, et al. (1996) Proc. Natl. Acad. Sci. USA 93:9816-9820; Ellingson, et al. (2005) J. Food Prot. 68:966-972; Grant, et al. (2002) Appl. Environ. Microbiol. 68:602-607; Ayele, et al. (2005) Appl. Environ. Microbiol. 71:1210-1214). There is increasing concern that MAP may be zoonotic (Greenstein & Collins (2004) Lancet 364:396-397; Greenstein (2003) Lancet Infect. Dis. 3:507-514; Greenstein, et al. (2009) In: Fratamico et al. (eds.) Sequelae and Long-Term Consequences of Infectious Diseases. One ed. Washington, ASM Press. American Society for Microbiology pp. 135-168). In contrast to leprosy, where M. leprae has never been grown in vitro (Stewart-Tull (1982) In: Ratledge & Stanford (eds.) The Biology of the Mycobacteria, Volume 1: Physiology, Identification, and Classification. One ed. New York, Academic Press. pp. 273-307), MAP has been cultured from humans with CD (Chiodini, et al. (1986) J. Clin. Microbiol. 24:357-363; Chiodini, et al. (1984) J. Clin. Microbiol. 20:966-971; Naser, et al. (2004) Lancet 364:1039-1044; Naser, et al. (2000) Am. J. Gastroenterol. 95:1094-1095; Bull, et al. (2003) J. Clin. Microbiol. 41:2915-2923) as well as patients with UC (aser, et al. (2004) supra).
There is an emerging explanation as to why this probable MAP zoonosis has not been appreciated. It is that multiple agents used in the treatment of IBD are in fact anti-MAP antibiotics. They are conventionally called “anti-inflammatories” (Greenstein, et al. (2007) PLoS ONE 2:e516) and “immuno modulators” (Shin & Collins (2008) Antimicrob. Agents Chemother. 52:418-426; Greenstein, et al. (2009) Int. J. Infect. Dis. 13:e254-263; Greenstein, et al. (2007) PLoS ONE 2:e161; Greenstein, et al. (2008) PLoS ONE 3:e249). Analogous to the multiple clinical manifestations of leprosy (Greenstein, et al. (2009) supra; Ridley & Jopling (1962) Lepr. Rev. 33:119-128; Ridley & Jopling (1966) Int. J. Lepr. Other Mycobact. Dis. 34:255-273), it has been suggested that all of IBD may be caused MAP (Mishina, et al. (1996) supra; Greenstein, et al. (2009) supra; Naser, et al. (2004) supra).
In many cases, the detection of MAP in living tissue, food, and water requires first culturing the bacterium. Even though MAP is hardy, it is slow growing and fastidious, which means it is difficult to culture. To facilitate detection of MAP, improved culturing methods are needed. The present invention addresses this need in the art by providing a culture medium which enhances the growth of MAP thereby facilitating its detection in food, water, and other biological samples.